1.Efficacy of topical application of PTD-SOD on wound healing in mice with mechanical injury
Yuangang LIU ; Yu CHENG ; Chen ZHANG ; Shutao LIU ; Pingfan RAO
Chinese Journal of Trauma 2011;27(6):566-572
Objective To study the role of the oxidative stress in the development of wound healing and observe the effect of the antioxidant PTD-SOD on damage and inflammation reaction after mechanical wound. Methods In this experiment,acute wound healing model by removal the whole layer dorsal skin of the mice was prepared,SOD(3 000 U and 6 000 U)and the fusion protein PTD-SOD with different concentrations(1 000 U,3 000 U,6 000 U and 10 000 U)were used to deal with the wounds continuously for 13 days.The mice were divided into different concentration SOD treatment group and PTD-SOD treatment group,model control group,physiological saline treatment group and compound iodine solution control group.The wound healing situation and healing percentage of the fight and left skin wounds of each mouse in every group was recorded every day.At day 14 after wound,the wound healing skin of each group was removed and some were used to make 10%tissues homogenate for detecting the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and contents of malondialdehyde(MDA)and hydroxyproline(Hyp);in the meantime,the other removed skin were fixed in 10% formalin for observing the histopathological changes of the tissues. Results Compared with the model control group,the physiological saline treatment group and the compound iodine solution control group,the skin wound healing percentage was significantly(P<0.05 or P<0.01)improved,with increase of the activities of SOD,CAT,GSH-Px and contents of Hyp (P<0.05 or P<0.01)and decrease of MDA(P<0.05 or P<0.01) in the SOD groups or PTD-SOD groups (except for 10 000 U PTD-SOD group).When compared with the physiological saline treatment group or the compound iodine solution treatment group,the effect was similar to the model control group.In comparison to the SOD groups,under the same dosage and environment condition,the PTD-SOD groups were much better than SOD groups with regard to promoting skin wound healing percentage,increasing activities of antioxidases and contents of Hyp,decreasing contents of MDA.Among the PTD-SOD groups,the effect of high dosage 10 000 U on promoting skin wound healing was declined. Conclusions The oxidative stress may playan important role in the development of wound healing.Proper application of treatment with antioxidants is a alternative strategy in the early stage of wound.PTD-SOD is able to prevent the oxidative stress damage,inhibit inflammatory infiltration and promote skin wound healing efficiently.
2.Relationship between the levels of tumor necrosis factor alpha, interleukin-1beta and interleukin 6 of bone tissue and osteoporosis in ovariectomized rats
Shutao ZHU ; Yang LIU ; Minghui ZHANG ; Xiao WANG
Chinese Journal of Tissue Engineering Research 2016;20(15):2206-2211
BACKGROUND:It has found that secretions of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and interleukin 6 (IL-6) are closely related to osteoporosis
OBJECTIVE:To further investigate the levels of TNF-α, IL-1β and IL-6 of bone tissue in ovariectomized rats and their relationship with osteoporosis.
METHODS: Sixty 3-month-old female Sprague-Dawley rats were randomly divided into control group and test group (n=30 per group). In the test group osteoporosis model was induced by ovariectomy folowed by intraperitoneal injection of aluminum nitride solution; while rats in the control group were given intraperitoneal injection of normal saline without ovariectomy. The experiment cycle was 12 weeks. One week after modeling, serum level of estradiol was tested, and the bone mineral density of rats measured with bone densitometry. In the meanwhile, the levels of TNF-α, IL-1β and IL-6 in bone tissue were detected to analyze the correlation between cytokines and bone mineral density.
RESULTS AND CONCLUSION: The level of serum estradiol in the test group was significantly lower than that in the control group (P=0.007); the bone mineral density of the lumbar spine and femur in the test group was significantly lower than that in the control group (P=0.006, 0.004). Compared with the control group, the percentages of trabecular bone and osteoblast area within the visual field in the test group were significantly lower, while the osteoclast area within the visual field was significantly higher (P=0.037, 0.029, 0.044). Compared with the control group, the levels of TNF-α, IL-1β and IL-6 in the test group significantly increased (P=0.032, 0.031, 0.025). And the bone mineral density of the lumbar spine and femur was negatively correlated with the levels of TNF-α, IL-1β and IL-6 in the test group (P < 0.05). In conclusion, the osteoclast activity is enhanced by elevating the levels of TNF-α, IL-1β and IL-6 of bone tissue of ovariectomized rats, and increasing the percentage of osteoclast area within the visual field. Moreover, the percentages of trabecular bone and osteoblast area within the visual field have a decline, which accelerates the process of bone resorption and leads to the formation of osteoporosis. Subject headings:Ovary; Cytokines; Osteoporosis; Tissue Engineering
3.A study of the blood supply relevant to prevention of nipple-areola necrosis in female breast operation
Dezong GAO ; Jingzhong SUN ; Qunsheng YIN ; Shutao LIU
Chinese Journal of General Surgery 1997;0(04):-
Objective To study the arterial blood supply of nipple-areola and provide the anatomical basis for avoiding nipple-areola necrosis in breast operation. Methods The vascular structure of nipple-areola of 26 female breasts in 13 cadavers were studied. Results The nipple-areola mainly accepted arterial blood supply from branches of the lateral thoracic artery and the internal thoracic artery. The 2nd~4th intercostal ~perforating branches of the internal thoracic artery and branches of the lateral thoracic artery reach the base of nipple-areola from a superior,medial and upper lateral direction by passing between lobules of mammary gland, then ascend between the lacteal ducts to supply the nipple-areola; the perforators of the lateral thoracic artery and the superticial breast perforators of internal thoracic artery, formed extensive anastomoses ~subcutaneously , and particulatly under areola formed arterial rete, from which branches were given out to ~nipple-areola . The intercostal perforators and thoracoacromial perforators did not supply the nipple-areola. Conclusions When nipple-sparing mastectomy is performed, in order to avoid nipple-areola necrosis,it is necessary to protect the arterial rete under the areola, and thus, the thickness of areolar skin flap should not be less than 0.5cm; to ensure the blood supply of nipple-areola from the internal thoracic artery and the ~lateral thoracic artery in breast reduction, the superior-medial or superior-lateral breast pedicle should be used and the thickness of preserved posterior breast should not be less than 1.5cm.
4.The role of microRNA-21 in esophageal squamous cell carcinoma and its effects on the PDCD4
Tao LIU ; Qing LIU ; Wenjing MA ; Tuersun AERZIGULI ; Shutao ZHENG ; Xiaomei LU
Chinese Journal of Digestion 2011;31(8):536-539
Objective To investigate the role of microRNA-21 (miRNA-21) in promoting proliferation of Kazakh's esophageal squamous cell carcinoma (ESCC) cell line Eca109 and Kazakh's ESCC tissues, and its effects on the expression of programmed cell death 4(PDCD4)gene.Methods TheEca109cellsweredividedintomiRNA-21Mimicsgroup(transfersequence:5′-UCAACAUCAGUCUGAUAAGCUA-3′),miRNA-21inhibitorgroup(transfersequence:5′-UAGCUUAUCAGACUGAUGUUGA-3′), negative control group (the transfer random sequence)and normal control group (no transfection).The cells were seeded at 4.5 × 105/well in 6-well cell culture plates. According to RNA interference technology,thecells were transfected with LipofectamineTM 2000. After transfection, the cell proliferation was determined by cell number counting.The expression of miRNA-21 was detected by real-time quantitative polymerase chain reaction (qRT-PCR), and the expression of PDCD4 protein was measured by Western blot.Meanwhile, the expression of miRNA-21 and PDCD4 protein were performed in 18 pairs of Kazakh's ESCC and corresponding adjacent normal esophageal mucosa.ResultsAt 48 hour after transfection,compared with normal control group, the cell number increased 36% (P=0.002) in miRNA-21 Mimics group, decreased 28% (P=0.002) in miRNA-21 inhibitor group, and did not change significantly in negative control group (P=0.515). At 48 hour after transfection, the relative expression of miRNA-21 in miRNA-21 Mimics group, miRNA-21 inhibitor group, negative control group and normal controlgroup was 0.37±0.10, 9.17± 1.08, 0.74±0.23 and 1.04±0.34,respectively, and the relative protein expression of PDCD4 was 1.47 ± 0.11, 0.61±0.09, 0.89 ±0.12 and 0.79±0.02 accordingly.Compared with normal control group, the relative expression of miRNA-21 decreased (P=0.031) and the relative protein expression of PDCD4 up regulated (P=0.001) in miRNA-21 inhibitor group, the relative expression of miRNA-21 increased (P=0.001) and the relative protein expression of PDCD4 down regulated (P=0.030) in miRNA-21 Mimics group,and there was no significant difference in negative control group (P=0.272 and 0.541).In 16 pairs of Kazakh's ESCC tissues, the expression of miRNA-21 was significantly higher in tumor tissues (0.11 ±0.09) than that of corresponding adjacent esophageal normal tissues (0.03 ± 0.03, P=0.001), while the relative protein expression of PDCD4 was significantly lower (0.92 ± 0.39) than that of corresponding adjacent normal esophageal tissues (1.57 ± 0.80, P=0.004). And the higher expression of miRNA-21, the lower relative protein expression of PDCD4 (r=-0.538, P=0.046).ConclusionmiRNA-21 may promoted the cell proliferation through inhibiting PDCD4 expression,which involved in the pathogenesis of Kazakh's ESCC.
5.The expression of microRNA let-7 in esophageal squamous cell carcinoma and its clinicopathological significance
Qing LIU ; Guodong LU ; Shutao ZHENG ; Tao LIU ; Sheyhidin ILYAR ; Renyong LIN ; Xiaomei LU
Chinese Journal of Digestion 2011;31(4):231-234
Objective To estimate the effect of microRNA (miRNA) let-7 expression on human esophageal squamous cell carcinoma(ESCC) and the relationship between let-7 level and clinicopathological parameters. Methods ESCC cell line (Eca109) was transfected with let-7 or its inhibitor by RNAi and cell transfection techniques. Normal cultured Eca109 cell was served as negative control. The proliferation of Eca109 cell was detected by MTT. The expression of let-7 in Eca109 cells and 45 paired ESCC tissues and corresponding para-cancerous tissues were measured using real-time quantitative polymerase chain reaction (qRT-PCR). The relationship between let-7 level and clinicopathological parameters in patients with ESCC was analyzed. Results The A value of let-7 in Eca109 cells transfected with let-7 was lower than negative control (P=0.005), while it was higher in Eca109 cells transfected inhibitor than that in negative control 72 hours after transfection. In comparison with negative control, the expression of let-7 in Eca109 cells transfected with let-7 was increased 33% (1.33 vs 1.00,P=0. 039) and it was decreased 50% in Eca109 cells transfected with inhibitor (0.50 vs 1.00,P=0. 014). The ratio of let-7 expression in ESCC tissue and para-cancerous tissue was 0.66 ± 0.47 with significant differece (P= 0.001). Moreover, The level of let-7 expression in Han patients with ESCC was lower than Kazakh patients with ESCC (0.48±0.43 vs 0. 88±0.51,P=0. 019). The level of let-7 expression in poorly differentiated ESCC tissue was lower than well differentiated ESCC tissue (0.42±0.30 vs 0.84±0.38,P=0. 015). The level of let-7 expression in patients with lymph node metastasis was lower than those without lymph node metastasis (0.50±0.35vs 0. 80±0.52,P=0. 032) . Conclusion It is demonstrated that let-7 can inhibit the carcinogenesis and development of ESCC. The level of let-7 expression is associated with cell differentiation,lymph node metastasis and nationalities.
6.The association analysis of catechol-O-methyltransferase gene Val158Met polymorphisms with esophageal squamous cell carcinoma in Yili Prefecture of Xinjiang
Conggai HUANG ; Tao LIU ; Guodong Lü ; Qing LIU ; Junguo FENG ; Shutao ZHENG ; Sheyhidin ILYAR ; Xiaomei LU
Chinese Journal of Digestion 2011;31(1):6-10
Objective To explore the relationship between polymorphism of catechol-O-methyltransferase (COMT) gene valine (Val) 158 methionine (Met) (G to A transition)and the distribution in population and esophageal squamous cell carcinoma (ESCC) in Yili prefecture of Xinjiang.Methods A hospital based case-control study was adopted, a total of 622 subjects, which including 214 ESCC patients and 408 age, gender and ethnicity-matched normal control individuals.The polymorphism of COMT gene G to A transition was analyzed with PCR-restriction fragment length polymorphism approaches.Results The COMT genotype frequencies in 622 subjects in Yili prefecture were GG genotype accounted for 47.3%, GA type for 42.3% and AA type for 10.4%, G allele was 68.4% and A allele was 31.6%.There was no statistical difference in the COMT genotype and frequencies of allele distribution between ESCC group and control group.Furthermore, stratified analysis indicated that there was statistical difference between ESCC group and control group in subjects less than 60 years old.There was statistical difference in the allele distribution among Kazak,Uygur and Han ESCC groups.The COMT genotype and frequency of allele distribution among normal control groups of the three ethnic groups were statistically different.After corrected age and gender,there was no statistical difference in COMT Val158Met polymorphisms among Kazakh, Uygur and Han ethnic groups in both ESCC and control groups in Yili Prefecture of Xinjiang.Conclusion COMT gene Val158Met single nucleotide polymorphism may not be the genetic markers of ESCC risk in Yili Prefecture of Xinjiang.
7.Effects of MMP-3 Gene Polymorphism in Restenosis after Percutaneous Coronary Interventions
Minli CHEN ; Jibing DU ; Yin LIU ; Jing GAO ; Rangzhuang CUI ; Shutao CHEN ; Hongliang CONG
Tianjin Medical Journal 2013;(11):1063-1066
Objective To investigate the relationship between matrix metalloproteinase-3 (MMP-3) gene promoter polymorphisms 5A/6A and the restenosis after percutaneous coronary intervention (PCI). Methods A total of 437 patients with PCI were selected in this study. Patients were divided into mutant genotype group (5A/5A+5A/6A, n=136) and wild genotype group (6A/6A, n=301) according to MMP-3 polymorphism. The angiography and clinic data were collected before and after coronary angiography in two groups of patients. The serum level MMP-3 and genotype analysis were compared be-tween two groups. Results There was no significant difference in the restenosis rate between two groups (42.2%vs 33.1%, P>0.05). The restenosis degree was significantly higher in wild genotype group than that in mutant genotype group (56.28%± 11.10%vs 36.00%±10.17%, P<0.01). There was no significant difference in the serum level of MMP-3 between two groups (13.38μg/L ± 3.00μg/L vs 12.33μg/L ± 2.96μg/L, P>0.05). There was a higher restenosis rate in patients carrying 6A al-lele than that of patients carrying 5A allele (P<0.05). Carrying wild genotypes are risk factors for restenosis after PCI. Con-clusion Patients carrying 6A allele have significantly higher risk of resteonsis than patients carrying 5A allele.
8.The studies on the changes of nitric oxide synthase in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the secondary lung injuris
Rui LI ; Jiangwei LIU ; Jianhui QIAN ; Ruoyun QIAN ; Qiong ZHANG ; Shutao ZHENG ; Fan YANG
Chinese Journal of Emergency Medicine 2015;24(2):163-168
Objective To study the pathological changes and expressions of NO and iNOS mRNA in the lung tissue of traumatic hemorrhagic shock rats under dry heat environment of desert and their relations to the lung injury.Methods A total of 140 male SD rats were randomly (random number) ivided into the room temperature (25 ℃) environment traumatic hemorrhagic shock group (room temperature group) and the dry heat traumatic hemorrhagic shock groups (dry heat group,temperature 40℃,humidity 10%),respectively,and each groups was further randomly divided into 7 subgroups:the control subgroup,post shock subgroups at 0,0.5,1,1.5,2and 3 h (n =10 in each subgroup).The rats of control subgroup were not treated,and rats of dry heat group were placed in dry heat environment for 60 min,then anesthetized,fixed,and insertion of intravenous indwelling needles and catherization of right carotid artery,jugular vein and the right femoral artery were performed.After stabilization for 10 min,2500 g iron wheel was used to be dropped from 30 m height and vertically hit the upper left femoral of SD rats in order to make comminuted fracture,wounds were quickly dressed after injury.Exsanguination from right femoral artery was kept until MAP maintained at (35 ± 5) mmHg,and resuscitation was carried out after continue monitoring for 60 min.After the establishment of traumatic hemorrhagic shock model in each environment,the rats were sacrificed at given intervals,and thoracotomy was performed to take broncho-alveolar lavage fluid (BALF) and lung tissue.Pathological changes of lung tissues were observed by using HE staining and NO concentration of lung tissue was detected by one-step method,and changes of the iNOS mRNA expressions were detected by using fluorescence quantitative PCR.Then t test,ANOVA and Pearson correlation analysis were used for the data analysis.Results The pathological change in dry heat group at each interval was more severe,and pulmonary histopathological injury score was higher,and the protein exudation was more profuse compared with the room temperature group.NO concentration in lung tissue homogenate of dry heat group was higher than that of room temperature group (t =2.472,P < 0.05),and the difference in NO level between different intervals within the dry heat group was statistically significant (F =6.77,P < 0.01).The NO concentration in dry heat group reached its maximum at 2 h (3.35 ± 0.23) μmol / g and the peak value emerged sooner than that in room temperature group.The difference was statistically significant in overall expression of iNOS mRNA between two groups analyzed with t test (t =3.619,P < 0.01),and there was statistically significant difference between intervals within the dry heat group (F =12.34,P <0.01).The values of iNOS mRNA in the dry heat group were higher than those in the room temperature group at the same given intervals,and the peak value appears at 1.5 h in dry heat group,and the room temperature group it began to increase at 2 h.The concentration of NO and the expression of iNOS mRNA were positively correlated with each other in two groups (r =0.680,r =0.376).The expression of iNOS mRNA and lung histopathological injury score was positively correlated in two groups (r =0.846,r =0.899).Conclusions When traumatic hemorrhagic shock occurred in the dry heat desert environment,the lung injury was more severe and appeared sooner than that in the room temperature environment.NO and iNOS played important roles in the secondary lung injury in the wake of traumatic hemorrhagic shock in rats under the dry heat environmengt of desert.
9.The Synchronous Detection and Significance of β-Glucuronidase Both in Serum and Tumorous Tissue in Non-Hodgkin Malignant Lymphoma Patient
Hong ZHANG ; Rui ZHENG ; Bo YANG ; Jing HU ; Bin XU ; Shutao HE ; Anmin YU ; Yingjie LIU
Journal of China Medical University 2001;30(1):4-5,30
Objective: Our purpose was to explore the change regularity of β-glucuronidase (β-G) in body of patients with Non-Hodgkin malignant lymphoma. Methods: β-G was examined synchronously both in the serum and in the tumor tissue of 13 cases patient with Non-Hodgkin malignant lymphoma by using the method of enzymlinked immunsorbent assay (ELISA) and immunohistochemistry separately. Among them, 3 cases were studied by using the immuno electron microscopic technique. Results: β-G was highly expressed both in the serum and tumorous tissue in patients with non-Hodgkin malignant lymphoma and there was obviously difference as compared with the control group (P<0.01). Conclusion: The combined detection with functional and morphological methods to β-G, it may be assistant target to early discovery and early diagnosis of Non-Hodskin malignant lymphoma.
10.Annexin A2 expression in human esophageal squamous cell carcinoma and correlation with invasion and metastasis
Junguo FENG ; Shutao ZHENG ; Hui LIU ; Tao LIU ; Conggai HUANG ; Qing LIU ; Renyong LIN ; Sheyhidin ILYAR ; Xiaomei LU
Chinese Journal of Digestion 2010;30(11):798-802
Objective To explore Annexin A2 expression in human esophageal squamous cell carcinoma (ESCC) and investigate the correlation of Annexin A2 expression with invasion and metastasis of human ESCC. Methods From 2000 to 2008, specimens of Xinjiang medical University First Affiliated Hospital were collected. Pathologically confirmed ESCC surgical specimens were set as experimental group, and the corresponding tumor adjacent tissues located more than 5 cm far from ESCC center were set as control group. 22 fresh and 175 paraffin-embeded ESCC specimens with corresponding adjacent tissues were randomly collected as study samples. With qRT-PCR, Western-blot and immunohistochemistry, the expression of Annexin A2 were detected at the mRNA and protein level. The correlation between Annexin A2 expression and clinicopathological parameters was analyzed. Results In 22 pairs of fresh ESCC and corresponding tumor adjacent tissues, the expression of Annexin A2 at mRNA level was significantly higher in tumor adjacent tissues (0. 06 ± 0. 06) than that in ESCC (0. 02 ±0. 02) (P<0.05 ). Annexin A2 expression at protein level was also significantly higher in tumor adjacent tissues (0.95±0. 42) than ESCC (0.81±0. 36) (P<0.05). In 175 paraffin-embeded ESCC specimens and corresponding adjacent tissues, the positive rate of Annexin A2 protein expression was 82. 3% (144/175) of the ESCC samples, which was lower than corresponding tumor adjacent tissues 92. 0% (161/175)(P<0. 05). In addition, Annexin A2 expression was correlated with lymphoid node metastasis (P<0.05) and pathological differentiation in patients with ESCC (P<0.05). However, there was no apparent correlation with gross type (P>0. 05). Conclusion The low expression of Annexin A2 in ESCC maybe played a potential role in the carcinogenesis, invasion and metastasis.